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Xiao-Xi Li,1,* Hong Li,2,* Li-Quan Jin,3 Yun-Bo Tan1,3 1Dali University of Clinical Medicine School, Dali, Yunnan, 671000, People’s Republic of China; 2Department of Radiology, Affiliated Renhe Hospital of China Three Gorges University, Hubei, 443001, People’s Republic of China; 3Department of General Surgery, The First of Affiliated Hospital of Dali University, Dali, Yunnan, 671000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yun-Bo Tan, Department of General Surgery, the First of Affiliated Hospital of Dali University, No. 32 of Carlsberg Road, Dali, Yunnan, 671000, People’s Republic of China, Tel +86 15517162885, Email tanyunbo6365@163.comObjective: To find pancreatic cancer (PC)-related hub genes based on weighted gene co-expression network analysis (WGCNA) construction and immune infiltration score analysis and validate them immunohistochemically by clinical cases, to generate new concepts or therapeutic targets for the early diagnosis and treatment of PC.Material and Methods: In this study, WGCNA and immune infiltration score were utilized to identify the relevant core modules of PC and the hub genes within these core modules.Results: Using WGCNA analysis, data from PC and normal pancreas integrated with TCGA and GTEX were analyzed and brown modules were chosen from the six modules. Five hub genes, including DPYD, FXYD6, MAP6, FAM110B, and ANK2, were discovered to have differential survival significance via validation tests utilizing survival analysis curves and the GEPIA database. The DPYD gene was the only gene associated with PC survival side effects. Validation of the Human Protein Atlas (HPA) database and immunohistochemical testing of clinical samples showed positive results for DPYD expression in PC.Conclusion: In this study, we identified DPYD, FXYD6, MAP6, FAM110B, and ANK2, as immune-related candidate markers for PC. Only the DPYD gene had a negative impact on the survival of PC patients. Through validation of the HPA database and immunohistochemical testing of clinical cases, we believe that the DPYD gene brings novel ideas and therapeutic targets in the diagnosis and treatment of PC.Keywords: DPYD, pancreatic cancer, tumor immunity, WGCNA

Background: To develop a laparoscopic training course that combines a smartphone application (APP) and virtual reality (VR), and initially evaluate the feasibility and effectiveness of its implementation., Methods: The Exploring Laparoscopy (Ex-Lap) app was developed to meet training demands. The course was designed by integrating the app with a VR simulator (LapSim ® ) and animal organ perfusion simulators. From January 2021 to December 2023, 91 participants were enrolled in the study and then divided into 5 separate batches to undergo the first stage of the course. The performance of the participants was evaluated by rating scale, the overall Training and Assessment of Basic Laparoscopic Techniques (TABLT) scores, and pass rates. Statistical analyses were conducted using SPSS 26.0, employing Kruskal-Wallis tests, Chi-squared analysis, and Fisher's exact test, depending on the data type., Results: The Staged Training and Assessment of Laparoscopic Skills (STALS) course was developed, consisting of three stages. The overall pass rates for the first stage across the five batches ranged from 85 to 100%, with no significant difference (P = 0.387). No significant differences were found in the scale scores or TABLT scores for the training tasks among students from different batches (all P > 0.05)., Conclusions: The STALS course is applicable in residency training, demonstrating satisfactory teaching effectiveness and replicability., (© 2024. The Author(s).)

Introduction: Ethnic differences associated with oral glucose tolerance test (OGTT) phenotypes is less studied in Southeast Asian ethnicities, especially in women with hyperglycemia in pregnancy (HIP)., Research Design and Methods: We retrospectively examined 3027 women at KK Women's and Children's Hospital in 2019. Of these, 508 (16.8%) women were diagnosed with HIP using the IADPSG (International Association of Diabetes and Pregnancy Study Groups) criteria at 24-28 weeks. OGTT phenotypes were classified into four mutually exclusive groups based on abnormal plasma glucose at (1) 0 hour only; (2) 1 hour only; (3) 2 hour only; (4) ≥2 timepoints (reference). Multinomial logistic regression was used to examine the association between ethnicity and OGTT phenotypes, adjusting for maternal age, parity, and first-trimester body mass index., Results: Overall HIP prevalence was 16.8%, highest among Indians (20.5%), then Chinese (18.3%) and Malays (14.2%). Indians (relative risk ratio (RRR) 3.05) and Chinese (RRR 2.33) were at higher risk of displaying a fasting-only phenotype compared with Malays. Chinese were at increased risk of displaying a 2-hour postprandial phenotype with an RRR of 2.88 as compared with Malays., Conclusions: Unique OGTT phenotypes exist across ethnic groups among women who developed HIP in a multi-ethnic Asian population., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Introduction: Individuals with psychiatric disorders tend to report having poorer bonds with their parents during their early years. These individuals often experience lower quality of life as well. This study investigated the associations between aspects of parental bonding and health-related quality of life (HRQOL) in a clinical sample of youths. It was hypothesised that high parental care and low parental overprotectiveness would be associated with higher levels of HRQOL., Methods: Data were obtained from a larger cross-sectional study. The sample consisted of 400 psychiatric outpatients: 191 patients aged 14-21 (mean ± standard deviation 18.1 ± 2.22) years and 209 patients aged 22-35 (28.0 ± 4.33) years. The Parental Bonding Instrument was used to measure parental care and overprotectiveness. Short Form-12 measured physical health (physical component summary [PCS]) and mental health (mental component summary [MCS]) components of HRQOL, and the 8-item Patient Health Questionnaire assessed depressive symptoms. These scales and a sociodemographic form were self-administered. Multivariable linear regression was used for analysis., Results: About half of the sample reported affectionless control for mothers (46.6%) and fathers (45.9%). After controlling for sociodemographic variables, no significant relationship was found between aspects of parental bonding and PCS scores. Maternal care was associated with MCS scores (β = 0.32, P < 0.01) and PHQ-8 scores (β = -0.12, P < 0.05)., Conclusion: Our results suggest that youths who experienced quality care from their mothers exhibit better mental health functioning despite their clinical diagnoses, which suggests that early maternal care exerts an overall long-term protective effect. Early parental education that promotes positive parenting practices could improve the overall HRQOL of individuals in adulthood despite their clinical diagnoses., (Copyright © 2024 Copyright: © 2024 Singapore Medical Journal.)

Introduction: Preconception is a critical period to optimise gamete function and early placental development, essential for successful conception and long-term maternal-child health. However, there is a lack of preconception services and consequently, global fertility rates continue to fall and mothers embark on their pregnancy journey in poor health. There is an urgent need to implement a holistic community-level preconception care programme to optimise risk factors for poor fecundability and improve long-term maternal-child health., Method: We reviewed current evidence on fecundability lifestyle risk factors, the efficacy of existing preconception interventions and the use of digital platforms for health optimisation, to create a new digital-based preconception intervention model that will be implemented via an app. We present the theory, content and mode of delivery of this holistic model targeting couples planning for pregnancy., Results: We propose a new model featuring a user-friendly mobile app, which enables couples to self-assess fecundability risks through a personalised risk score that drives a tailored management plan. This tiered management provides anticipatory guidance supported by evidence-based recommen-dations, and promotes ongoing engagement for behavioural optimisation and specialist referrals as required. Based on the health belief model, this new model delivered with a mobile app seeks to shift couples' perceptions about their susceptibility and severity of subfertility, benefits of making a change and barriers to change., Conclusion: Our proposed digital-based intervention model via a mobile app stands to enhance preconcep-tion care by providing personalised risk assessments, real-time feedback and tiered management to optimise preconception reproductive health of couples. This model forms a reference content framework for future preconception care intervention delivery., Competing Interests: This research was supported by the KKH Health Services Model of Care Transformation Fund (MoCTF) Grant (MoCTF/01/2020, MoCTF/02/2020 and MoCTF/03/2020) and the Lien Foundation Optimising Maternal and Child Health Programme Fund. The funding body did not influence either the data collection and analysis or the writing and the decision to submit the manuscript. CWK and JKYC are supported by the National Medical Research Council, Ministry of Health, Singapore (NMRC/MOH-000596-00 and NMRC/CSASI-008-2016, MOH-001266-01, MOH-001221-01 and MOH-000932-01, respectively). KMG is supported by the National Institute for Health Research (NIHR Senior Investigator (NF-SI-0515-10042), NIHR Southampton Biomedical Research Centre (IS-BRC-1215-20004), British Heart Foundation (RG/15/17/3174) and the European Union (Erasmus+ Programme ImpENSA 598488-EPP-1-2018-1-DE-EPPKA2-CBHE-JP).

Background: The COVID-19 pandemic brought on a range of stressors in homes and workplaces. With no sign of ending after one year, burnout was a concern. Resilience has been known to shield against the effects of stress. While often thought of as an individual trait, previous studies have shown social support to improve resilience. The study aimed to examine the extent of burnout in the Singapore population and whether social support and resilience cushioned the impact of COVID-19 related stressors a year into the pandemic., Methods: Participants were 858 Singapore residents who participated in a larger study between October 2021 and September 2022. The Copenhagen Burnout Inventory provided Work-and Personal-related burnout scores. Multivariable linear regression was used to identify demographic variables associated with burnout. Path analysis revealed the associations between COVID-19 stressors, social support, resilience and burnout., Results: 22 and 19% of the sample reported work and personal burnout respectively, with younger adults being more burnt out than older adults. Higher stress was associated with higher burnout and higher social support was associated with lower burnout. Path analysis revealed that the relationship between social support and burnout was partially accounted for by increased resilience., Conclusion: Managing altered work arrangements, career expectations, and increased responsibilities at home may contribute to greater levels of burnout in the younger adults. Increased employer support and targeted interventions could mitigate the impact of these stressors. The study also highlighted the importance of maintaining social connections even while physically distancing., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Shahwan, Tay, Shafie, Tan, Gunasekaran, Tan, Satghare, Zhang, Wang, Tan and Subramaniam.)

Introduction: This systematic review is aimed at (1) evaluating the association between media portrayals of suicides and subsequent copycat suicides or attempts among the general public in Asia, (2) understanding the factors associated with copycat suicides and (3) determining the positive impacts of the media reporting of suicides (e.g. increased help-seeking, coping)., Method: A systematic review and narrative synthesis of English and Chinese articles from 8 electronic databases (i.e. PsycINFO, MEDLINE, Embase, CINAHL, Web of Science, Ariti, China National Knowledge Infrastructure and OpenGrey) from January 2000 to May 2023 was conducted. Observational studies were included, and the data were analysed through narrative synthesis. The protocol was registered with PROSPERO (CRD42021281535)., Results: Among the 32 studies included (n=29 for evidence synthesis) in the review, there is good-quality evidence to show that copycat suicides and suicide attempts increase after media reports of a suicide, regardless of country, celebrity status, study design, type of media, mode of suicide or follow-up period. Females, younger age groups and those sharing similar characteristics as the deceased in publicised suicides (age, gender) were more susceptible to negative impact. Reporting of the mode of death of the deceased increased suicides by the same method among the public., Conclusion: Media portrayals of suicide appear to have a negative impact on copycat suicides at the population level in Asia. Thus, in addition to tighter media control, healthcare systems, professional medical bodies and community outreach services should work collaboratively to promote early help-seeking in those with psychological distress.

Background: Open-access oesophagogastroduodenoscopy (OAO) is defined as the performance of oesophagogastroduodenoscopy (OGD) requested by referring physicians without a prior specialist consultation. With the increasing demand for specialist appointments, the use of OAO has helped to reduce healthcare utilization by decreasing prior clinic visits. This also allows endoscopies to be scheduled and performed earlier. This study aims to evaluate our experience in providing OAO services to patients with non-alarming dyspepsia symptoms under the age of 60., Methods: The records of patients scheduled for OAO from January 2019 to December 2022 at Singapore General Hospital (SGH) Department of Gastroenterology were analyzed., Results: Five hundred sixty-nine patients were scheduled for OAO, and 436 patients underwent the procedure. The mean age of patients was 45.7 (SD=10.9) years old. Thirty-six percent were males, and there were 80.8% Chinese, 5.3% Malay, 8.6% Indian, and 5.3% others. The median waiting time for endoscopy was 23 days (IQR 16-36), and no major adverse events were reported. Over half of the endoscopies were unremarkable (n=231, 53%). There were 25 (5.7%) patients with major findings; three had upper gastrointestinal adenocarcinoma (one oesophageal and two gastric), one had oesophageal varices, and 21 had peptic ulcer disease (10 gastric and 11 duodenal ulcers). A rapid urease test was conducted on 409 patients, and 55 (13.4%) were positive., Conclusion: OAO is a safe and effective strategy for providing timely diagnostic OGD to normal-risk patients at our center. Primary care physicians are encouraged to refer non-alarming dyspepsia symptoms patients under 60 years for OAO over the conventional route., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Tan et al.)

Background and Aim: Colonoscopy is commonly used in screening and surveillance for colorectal cancer. Multiple different guidelines provide recommendations on the interval between colonoscopies. This can be challenging for non-specialist healthcare providers to navigate. Large language models like ChatGPT are a potential tool for parsing patient histories and providing advice. However, the standard GPT model is not designed for medical use and can hallucinate. One way to overcome these challenges is to provide contextual information with medical guidelines to help the model respond accurately to queries. Our study compares the standard GPT4 against a contextualized model provided with relevant screening guidelines. We evaluated whether the models could provide correct advice for screening and surveillance intervals for colonoscopy., Methods: Relevant guidelines pertaining to colorectal cancer screening and surveillance were formulated into a knowledge base for GPT. We tested 62 example case scenarios (three times each) on standard GPT4 and on a contextualized model with the knowledge base., Results: The contextualized GPT4 model outperformed the standard GPT4 in all domains. No high-risk features were missed, and only two cases had hallucination of additional high-risk features. A correct interval to colonoscopy was provided in the majority of cases. Guidelines were appropriately cited in almost all cases., Conclusions: A contextualized GPT4 model could identify high-risk features and quote appropriate guidelines without significant hallucination. It gave a correct interval to the next colonoscopy in the majority of cases. This provides proof of concept that ChatGPT with appropriate refinement can serve as an accurate physician assistant., (© 2023 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.)

Analyzing the pattern of altitudinal variation in the leaf traits and their networks of a particular tree species of similar age and its influencing factors could contribute to understanding the impacts of environmental factors on leaf traits and excluding the interference of genetic factors. We investigated the stomatal, structural, chemical, and vein traits of Daphniphyllum macropodum leaves in middle-aged forests, following the altitudinal gradient (1100, 1500, and 1900 m) on Mao'er Mountain. The objectives of this study were to reveal patterns in leaf trait and leaf trait networks variation, the life strategy of the tree species, and the major environmental factors affecting the altitudinal variations. The results showed that leaf area, specific leaf area, leaf thickness, leaf dry matter content, chlorophyll content, nitrogen content, phosphorus content, C:N, C:P, vein density, and vein diameter varied significantly across altitudes. Mean annual temperature and total radiation explained 42.1% and 16.2% of leaf-trait variation, respectively. They served as key environmental factors driving the altitudinal variation in leaf traits. Mean annual temperature exhibited the greatest influence on leaf area ( R 2 =0.73), and total radiation exerted the most prominent effect on leaf thickness ( R 2 =0.72). Both relationships were significantly positive. D. macropodum exhibited low leaf nitrogen and phosphorus at the low altitude of 1100 m, and the overall and local trait networks were loose, adopting a conservative resource strategy. At the medium altitude of 1500 m, leaf nutrient contents were relatively high. The overall network of leaf traits was tightly connected and local network was loose. By enhancing the dependency among leaf traits, and improving phosphorus utilization efficiency, D. macropodum could cope with competition in deciduous forests and adopt resource acquisition strategies. Further, at the highest altitude of 1900 m, D. macropodum had relatively large leaf thickness, chlorophyll content, and leaf dry matter content, but relatively small leaf area. The local network connections were tight while the overall network looseness, indicating a resource conserving strategy. The trade-off relationship between C:P and leaf phosphorus content was closely related to phosphorus use efficiency, and its variation was an important indicator for identifying life strategies of D. macropodum in different altitudes.

Chikungunya virus (CHIKV) nonstructural protein 1 (nsP1) contains both the N7-guanine methyltransferase and guanylyltransferase activities and catalyzes the 5' end cap formation of viral RNAs. To further understand its catalytic activity and role in virus-host interaction, we demonstrate that purified recombinant CHIKV nsP1 can reverse the guanylyl transfer reaction and remove the m 7 GMP from a variety of capped RNA substrates including host mRNAs. We then provide the structural basis of this function with a high-resolution cryo-EM structure of nsP1 in complex with the unconventional cap-1 substrate RNA m 7 GpppA m U. We show that the 5'ppRNA species generated by decapping can trigger retinoic acid-inducible gene I-mediated interferon response. We further demonstrate that the decapping activity is conserved among the alphaviral nsP1s. To our knowledge, this is a new mechanism through which alphaviruses activate the antiviral immune response. This decapping activity could promote cellular mRNA degradation and facilitate viral gene expression, which is functionally analogous to the cap-snatching mechanism by influenza virus., Competing Interests: Conflict of interest The authors declare that they have no conflict interests with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Increasing research efforts have focused on understanding why some individuals develop severe psychopathology after exposure to adverse childhood experiences (ACEs). Sleep disturbances (insomnia, nightmares, and sleep disorders) are prevalent sequelae of ACEs and associated with psychopathology; however, there is no meta-analytic evidence on whether sleep disturbance functions as a transdiagnostic mediator in the relationship between ACEs and psychopathology (internalizing/externalizing disorders and psychosis) in children and adolescents., Methods: Systematic searches in three databases (PubMed; PsycINFO; Web of Science) identified 98 articles ( N  = 402,718; age range 1-17 years) and the present study used a novel two-stage meta-analytic structural equation model to investigate whether ACEs predict psychopathology through sleep disturbance. Subgroup analyses determined potential biases due to study design (cross-sectional vs. longitudinal) and geographical differences (Western vs. non-Western countries). Sensitivity analyses evaluated the influence of early childhood (<5 years old) and overlapping symptoms (i.e., nightmares and trauma symptoms) on model stability., Results: The pooled correlations among ACEs, sleep disturbance, and psychopathology were significant; the effect sizes ranged from moderate to high ( r  = 0.21 to r  = 0.29). The indirect effect from ACEs via sleep disturbance to psychopathology was significant ( β  = 0.05, 95% CI [0.04, 0.06]). The direct effect of ACEs on psychopathology was significant ( β  = 0.18, 95% CI 0.13-0.24). Subgroup analyses revealed larger effects for cross-sectional studies than longitudinal studies (Δ χ 2 (3) = 9.71, p  = 0.021). Sensitivity analyses revealed stable and consistent results., Conclusions: The present meta-analytic results indicate that sleep disturbance is a transdiagnostic mediator in the relationship between ACEs and psychopathology among children and adolescents. Further research is required to determine the synergistic effects between sleep disturbance and other risk mechanisms, and elucidate the complex pathways that lead to disorder in the aftermath of childhood adversities., Competing Interests: The authors declared that they have no competing or potential conflicts of interest., (© 2023 The Authors. JCPP Advances published by John Wiley & Sons Ltd on behalf of Association for Child and Adolescent Mental Health.)

Background and Aim: Endoscopic sleeve gastroplasty (ESG) is an effective treatment for obesity. Recently, a novel single-channel endoscopic suturing device has been made available to overcome the need for a double-channel endoscope. However, there is limited evidence evaluating its utility for ESG. In this multicenter study, we aim to assess the efficacy and safety of the single-channel suturing device for ESG., Methods: We reviewed the records of 18 patients who underwent ESG using the novel device at the Singapore General Hospital, Singapore, and Siriraj Hospital, Bangkok, between 2020-2021. We adopted a "U" suture pattern. Our primary outcome was to assess technical feasibility and safety. The secondary outcome was to determine the percentage of total body weight loss at 1 year., Results: The mean ± SD age and body mass index were 42 ± 8.5 years and 34.9 ± 4.4 kg/m 2 , respectively. The majority were female (61%). ESG was technically successful in 94% ( n  = 17) of patients. Device dislodgement occurred in one patient. We used an average of five sutures (range, 4-8), and the mean ± SD procedure time was 96.5 ± 43.8 min. No complications occurred. The mean ± SD length of stay was 2.3 ± 1.5 days. The mean ± SD percentage of total body weight loss at 6 and 12 months were 16 ± 5.2% and 13.1 ± 5.8%, respectively. We found that >5%, >10%, and >15% total body weight loss was observed in 83.3%, 72.2%, and 56%, respectively., Conclusion: ESG using the single-channel endoscopic suturing system is safe and effective for inducing weight loss at 1 year in patients with obesity., Competing Interests: Dr. Ravishankar Asokkumar is a consultant for Apollo Endosurgery, USA. The rest of the authors have no conflict of interest., (© 2023 The Authors. DEN Open published by John Wiley & Sons Australia, Ltd on behalf of Japan Gastroenterological Endoscopy Society.)

Many alphaviruses, including chikungunya virus (CHIKV) are known human pathogens that lack specific and effective antivirals or vaccines available. The upstream portion of the positive-sense single-stranded RNA genome of alphaviruses encodes four nonstructural proteins: nsP1 to nsP4. They are expressed and autoprocessed to nonstructural proteins which assemble into a replication complex (RC) playing multiple essential roles on viral RNA replication and communication with the host components. The assembly of alphavirus RC and its RNA genome initiates the membrane-derived ultrastructure known as spherule which facilitates viral RNA synthesis protected from host immune responses. Recent advances in the molecular understanding of the high-resolution CHIKV RC heteromeric ultrastructure have provided new insights into the viral replication process. Hence, alphavirus RC presents as an ideal multi-enzyme target for the development of structure-based antiviral drugs. Moreover, the alphavirus RC has therapeutic potential in the form of self-amplifying RNA technology against both infectious and non-infectious diseases., Competing Interests: Declaration of competing interest The authors declare that they have no competing interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This work reports (a) conglomerate and racemic crystal structures of [(Δ,Δ,Δ,Δ,Δ,Δ)- or/and (Λ,Λ,Λ,Λ,Λ,Λ)-Eu III 6 (TTP) 8 (OH 2 ) 6 Na 4 ] n coordination polymers, (b) racemic crystal structures of (Δ,Δ,Δ,Δ)-/(Λ,Λ,Λ,Λ)-Eu III 4 (TTP) 4 (bipy) 4 (MEK) 2 (OH 2 ) 2 tetrahedral clusters, and (c) the achiral crystal structure of the [Eu III 2 (BTP) 4 (OH 2 ) 2 Na 2 ] n coordination polymer (where BTP = dianionic bis-β-diketonate, TTP = trianionic tris- β -diketonate, and bipy = 2,2'-bipyridine). The screw coordination arrangement of the TTP ligand has led to the formation of homoconfigurational racemic Eu III products. The conglomerate crystallization of [Eu III 6 (TTP) 8 (OH 2 ) 6 Na 4 ] n appears to be caused by the presence of the sodium, Na + counterions, and interactions between oxygen atoms and the trifluoromethyl unit of the TTP ligand and Na + ions. All the Eu III compounds exhibit characteristic red luminescence ( 5 D 0 → 7 F J , J = 0-4) in solution or in the solid crystalline state. Circularly polarized luminescence (CPL) was observed in the chiral Eu III 6 (TTP) 8 (OH 2 ) 6 Na 4 ] n species, displaying a | g lum | value in the range of 0.15 to 0.68 at the 5 D 0 → 7 F 1 emission band. Subtle changes of the [Eu III 6 (TTP) 8 (OH 2 ) 6 Na 4 ] n structure which may be due to selection of twinned crystals or crystals that do not correspond to a perfect spontaneous resolution, are considered to be responsible for the variation in the observed CPL values., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

To better understand how positive-strand (+) RNA viruses assemble membrane-associated replication complexes (RCs) to synthesize, process, and transport viral RNA in virus-infected cells, we determined both the high-resolution structure of the core RNA replicase of chikungunya virus and the native RC architecture in its cellular context at subnanometer resolution, using in vitro reconstitution and in situ electron cryotomography, respectively. Within the core RNA replicase, the viral polymerase nsP4, which is in complex with nsP2 helicase-protease, sits in the central pore of the membrane-anchored nsP1 RNA-capping ring. The addition of a large cytoplasmic ring next to the C terminus of nsP1 forms the holo-RNA-RC as observed at the neck of spherules formed in virus-infected cells. These results represent a major conceptual advance in elucidating the molecular mechanisms of RNA virus replication and the principles underlying the molecular architecture of RCs, likely to be shared with many pathogenic (+) RNA viruses.

Starch is an ideal wall material for controlled release in oral delivery systems due to its non-allergic properties, availability, and cheap price. However, because of its poor mechanical behavior and high water permeability, it is necessary to modify the amphiphilic nature of starch. Surfactants are essential components to emulsify the lyophobic food ingredients. However, the interaction of starch with emulsifiers and how they affect the pasting behavior and digestion of starch are not well understood. In this paper, surfactants, such as non-ionic Tween (TW) and ionic sodium fatty acid (NaFA), with varying hydrophobic carbon chain lengths, were selected as model amphiphiles to investigate the structural, pasting, rheological properties and in vitro digestibility of regular and frozen starch samples. The results showed that, in most cases, the addition of TW reduced the viscosity of starch. However, saturated medium-chain NaFA increased the starch viscosity and rheological modulus greatly. Both surfactants inhibited starch digestion. This paper presents a comparative investigation on the effect of ionic and non-ionic surfactant on the structure and properties of corn starch, and therefore the information is useful for structural-based formulation with starch for developing colloidal delivery systems. It is also helpful for developing functional food with controllable digestion properties.

Introduction: Arthroscopic Bankart repair is a widely accepted procedure to treat recurrent shoulder dislocation. This study aims to describe our experience with arthroscopic Bankart repair and its functional outcome., Methods: 107 patients who underwent arthroscopic Bankart repair from 2008 to 2013 were followed up for a minimum of three years and reviewed by an independent observer. 80 consented to being interviewed using the Oxford Shoulder Instability Score (OSIS) and Simple Shoulder Test., Results: 82 shoulders (two bilateral) were studied. Mean age at first dislocation was 19.4 ± 3.4 (12.0-31.0) years. Mean follow-up was 4.4 ± 1.3 (3.0-9.0) years and 2.5 ± 3.0 (0.1-15.4) years elapsed from first dislocation to surgery. 41 (50.0%) patients played overhead or contact sports and 44 (53.7%) played competitive sports before injury; 8 (9.8%) patients reported recurrence of dislocation, which was significantly associated with playing competitive sports before injury (p <0.039), 5 (6.1%) underwent revision surgery and 22 (26.8%) reported residual instability after surgery. 49 (59.8%) patients returned to playing sports, 75 (91.5%) were satisfied with their surgery and 79 (96.3%) were willing to undergo the surgery again. 74 (90.2%) patients had two-year good/excellent OSIS, which was significantly associated with playing competitive sports before injury (p = 0.039), self-reported stability after surgery (p = 0.017), satisfaction with surgery (p = 0.018) and willingness to undergo surgery again (p = 0.024)., Conclusion: Arthroscopic Bankart repair yields good functional outcomes and is associated with high patient satisfaction, although not all patients return to sports., Competing Interests: None

Many viruses encode RNA-modifying enzymes to edit the 5' end of viral RNA to mimic the cellular mRNA for effective protein translation, genome replication, and evasion of the host defense mechanisms. Alphavirus nsP1 synthesizes the 5' end Cap-0 structure of viral RNAs. However, the molecular basis of the capping process remains unclear. We determine high-resolution cryoelectron microscopy (cryo-EM) structures of Chikungunya virus nsP1 in complex with m7GTP/SAH, covalently attached m7GMP, and Cap-0 viral RNA. These structures reveal details of viral-RNA-capping reactions and uncover a sequence-specific virus RNA-recognition pattern that, in turn, regulates viral-RNA-capping efficiency to ensure optimal genome replication and subgenomic RNA transcription. This sequence-specific enzyme-RNA pairing is conserved across all alphaviruses., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Alphaviruses such as Ross River virus (RRV), chikungunya virus (CHIKV), Sindbis virus (SINV), and Venezuelan equine encephalitis virus (VEEV) are mosquito-borne pathogens that can cause arthritis or encephalitis diseases. Nonstructural protein 4 (nsP4) of alphaviruses possesses RNA-dependent RNA polymerase (RdRp) activity essential for viral RNA replication. No 3D structure has been available for nsP4 of any alphaviruses despite its importance for understanding alphaviral RNA replication and for the design of antiviral drugs. Here, we report crystal structures of the RdRp domain of nsP4 from both RRV and SINV determined at resolutions of 2.6 Å and 1.9 Å. The structure of the alphavirus RdRp domain appears most closely related to RdRps from pestiviruses, noroviruses, and picornaviruses. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) methods showed that in solution, nsP4 is highly dynamic with an intrinsically disordered N-terminal domain. Both full-length nsP4 and the RdRp domain were capable to catalyze RNA polymerization. Structure-guided mutagenesis using a trans-replicase system identified nsP4 regions critical for viral RNA replication., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Background: Recent work suggests that APOEɛ4/4 females with Alzheimer's disease (AD) are more susceptible to developing neuropsychiatric symptoms (NPS)., Objective: To examine the interaction of sex and APOEɛ4 status on NPS burden using two independent cohorts: 1) patients at risk for AD with mild cognitive impairment and/or major depressive disorder (n = 252) and 2) patients with probable AD (n = 7,261)., Methods: Regression models examined the interactive effects of sex and APOEɛ4 on the number of NPS experienced and NPS Severity. APOEɛ3/4 and APOEɛ4/4 were pooled in the at-risk cohort due to the sample size., Results: In the at-risk cohort, there was a significant sex*APOEɛ4 interaction (p = 0.007) such that the association of APOEɛ4 with NPS was greater in females than in males (incident rate ratio (IRR) = 2.0). APOEɛ4/4 females had the most NPS (mean = 1.9) and the highest severity scores (mean = 3.5) of any subgroup. In the clinical cohort, APOEɛ4/4 females had significantly more NPS (IRR = 1.1, p = 0.001, mean = 3.1) and higher severity scores (b = 0.31, p = 0.015, mean = 3.7) than APOEɛ3/3 females (meanNPS = 2.9, meanSeverity = 3.3). No association was found in males., Conclusion: Our study suggests that sex modifies the association of APOEɛ4 on NPS burden. APOEɛ4/4 females may be particularly susceptible to increased NPS burden among individuals with AD and among individuals at risk for AD. Further investigation into the mechanisms behind these associations are needed.

The study of artists with acquired brain damage is an empirical way to investigate the multiplicity of cerebral changes that occur with artistic training. We describe a talented painter with a left progressive cerebral lesion. In spite of losing function of her right hand, she regained dexterity of the left one in ten days for painting and drawing but not for writing. We discuss two potential explanations for her rapid recovery: (a) her extensive artistic training and/or (b) the slow-growing nature of her cerebral lesion.

Objective: There is debate on the best method of colorectal cancer (CRC) surveillance in inflammatory bowel disease (IBD). We aimed to examine how gastroenterologists around the world practice CRC surveillance and manage dysplastic lesions in IBD., Methods: A 22-question survey was emailed to gastroenterologists from 34 countries. It included questions on resources for, frequency and mode of CRC surveillance, and management of colorectal dysplasia. Fisher's exact test and logistic regression were used to evaluate the differences among respondents in various domains., Results: There were 217 eligible responses, with most gastroenterologists working in public hospitals (76%), and treating >10 patients with IBD weekly (71%). High-definition white light endoscopy (HDWLE) was available in 93.1% of the centers. The preferred mode of surveillance was HDWLE with dye-spray chromoendoscopy and targeted biopsies (41.2%). Fewer than 50% of physicians reported using chromoendoscopy in >50% of cases, citing time as the limiting factor (73.7%). Of these gastroenterologists 63% infrequently (<25% of cases) performed random biopsies during chromoendoscopy. They would attempt endoscopic mucosal resection for polypoid lesions >10 mm (67.2%), including >20 mm lesions with low grade dysplasia (49.8%), and non-polypoid lesions >10 mm without dysplasia (56.9%). For non-polypoid lesions >20 mm with low- and high-grade dysplasia, referral to expert endoscopists was the preferred option., Conclusion: The preferred method of CRC surveillance was HDWLE with chromoendoscopy and targeted biopsies. Random biopsies were infrequently performed. The uptake of chromoendoscopy for surveillance in practice was low. Physicians varied in their approach in removing endoscopically resectable dysplastic lesions., (© 2021 Chinese Medical Association Shanghai Branch, Chinese Society of Gastroenterology, Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine and John Wiley & Sons Australia, Ltd.)

Chikungunya virus (CHIKV) causes a debilitating arthralgic inflammatory disease in humans. The multifunctional CHIKV protein, nsP1, facilitates virus RNA replication and transcription by anchoring the viral replication complex (RC) to plasma membrane vesicles and synthesizing the viral RNA 5' cap-0. Here, we report a cryo-EM structure of CHIKV nsP1 at 2.38 Å resolution. Twelve copies of nsP1 form a crown-shaped ring structure with a 7.5-nm-wide channel for mediating communication and exchange between the viral RC and the host cell. The catalytic site for viral RNA capping is located in a tunnel that is shaped by neighboring nsP1 molecules. Two membrane-association loops target nsP1 to the inner leaflet of the plasma membrane via palmitoylation and hydrophobic and electrostatic interactions. Our study provides the structural basis of viral RNA capping and RC assembly mediated by nsP1 and guides the development of antivirals targeting these essential steps of virus infection., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background and Aim: Real-world data on sofosbuvir/velpatasvir with and without ribavirin (SOF/VEL ± RBV), particularly among patients with genotype 3 (GT3) decompensated cirrhosis, prior treatment, coinfection, and hepatocellular carcinoma (HCC), are scarce. We aimed to assess the efficacy and safety of SOF/VEL ± RBV in a real-world setting that included both community and incarcerated GT3 hepatitis C virus (HCV) patients., Methods: We included all GT3 HCV patients treated with SOF/VEL ± RBV in our institution. The primary outcome measure was the overall sustained virological response 12 weeks after treatment (SVR12), reported in both intention-to-treat (ITT) and per-protocol analyses. The secondary outcome measures were SVR12 stratified by the presence of decompensated cirrhosis, prior treatment, HCC, and HIV/hepatitis C virus coinfection and the occurrence rate of serious adverse events requiring treatment cessation or hospitalization., Results: A total of 779 HCV patients were treated with 12 weeks of SOF/VEL ± RBV, of which 85% were treated during incarceration. Among the 530 GT3 HCV patients, 31% had liver cirrhosis, and 6% were treatment-experienced. The overall SVR12 for GT3 was 98.7% (95% confidence interval: 97.3%, 99.5%) and 99.2% (95% confidence interval: 98.1%, 99.8%) in ITT and per-protocol analyses, respectively. High SVR12 was also seen in ITT analysis among GT3 HCV patients with decompensated cirrhosis (88%), prior treatment (100%), HCC (100%), and HIV/hepatitis B virus coinfection (100%). Apart from one patient who developed myositis, no other serious adverse events were observed., Conclusion: The SOF/VEL ± RBV is a safe and efficacious treatment option for GT3 HCV patients in a real-world setting. SOF/VEL with RBV may be considered for decompensated GT3 HCV patients., (© 2020 Journal of Gastroenterology and Hepatology Foundation and John Wiley & Sons Australia, Ltd.)

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus responsible for chikungunya fever. Nonstructural protein 2 (nsP2), a multifunctional protein essential for viral replication, has an N-terminal helicase region (nsP2h), which has both nucleotide triphosphatase and RNA triphosphatase activities, as well as a C-terminal cysteine protease region (nsP2p), which is responsible for nonstructural polyprotein processing. The two functional units are connected through a linker of 14 residues. Although crystal structures of the helicase and protease regions of CHIKV nsP2 have been solved separately, the conformational arrangement of the full-length nsP2 and the biological role of the linker remain elusive. Using the small-angle X-ray scattering (SAXS) method, we demonstrated that the full-length nsP2 is elongated and partially folded in solution. The reconstructed model of the structure of nsP2 contains a flexible interdomain linker, and there is no direct interaction between the two structured regions. To examine the function of the interdomain linker, we constructed and characterized a set of CHIKV mutants. The deletion of three or five amino acid residues in the linker region resulted in a modest defect in viral RNA replication and transcription but completely abolished viral infectivity. In contrast, increasing the flexibility of nsP2 by lengthening the interdomain linker increased both genomic RNA replication and viral infectivity. The enzymatic activities of the corresponding mutant proteins were largely unaffected. This work suggests that increasing the interdomain flexibility of nsP2 could facilitate the assembly of the replication complex (RC) with increased efficiency and promote virus production. IMPORTANCE CHIKV nsP2 plays multiple roles in viral RNA replication and virus-host interactions. The helicase and protease regions of nsP2 are connected through a short linker. Here, we determined that the conformation of full-length CHIKV nsP2 is elongated and that the protein is flexible in solution. We also highlight the importance of the flexibility of the interdomain of nsP2 on viral RNA synthesis and infectivity. CHIKV mutants harboring shortened linkers fail to produce infectious virus particles despite showing only relatively mild defects in genomic and subgenomic RNA synthesis. Mutations increasing the length of the interdomain linker have only mild and generally beneficial impacts on virus replication. Thus, our findings link interdomain flexibility with the regulation of viral RNA replication and infectivity of the viral genome., (Copyright © 2021 American Society for Microbiology.)

Background: Gender differences in valvular heart disease are increasingly recognised. A prior study has suggested better surgical outcomes in women with symptomatic aortic stenosis (AS). We investigate gender differences in medically managed severe AS., Method: We studied 347 patients with severe AS (aortic valve area index <0.6 cm 2 /m 2 ) in terms of baseline clinical background, echocardiographic characteristics, and clinical outcomes. Appropriate univariate and multivariate models were employed, while Kaplan-Meier curves were constructed to compare mortality outcomes., Results: In total, 205 (59%) patients were women. Despite higher incidences of hypertension (75.6% vs 47.3%) and diabetes mellitus (46.5% vs 29.5%) in women, women had improved survival (Kaplan-Meier log-rank = 6.24, p = 0.012). After adjusting for age (hazard ratio [HR], 1.034; 95% confidence interval [CI], 1.014-1.054), hypertension (HR, 1.469; 95% CI, 0.807-2.673), diabetes (HR, 1.219; 95% CI, 0.693-2.145), and indexed aortic valve area (HR 0.145, 95% CI 0.031-0.684) on multivariate analyses, female gender remained independently associated with lower mortality (HR, 0.561; 95%, CI 0.332-0.947). Women tended to have smaller body surface area (BSA), left ventricular (LV) internal diastolic diameter, and smaller LV outflow tract diameter but were similar to men in terms of LV ejection fraction, AS severity, and patterns of LV remodelling., Conclusions: Women appeared to have better outcomes compared to men in medically managed severe AS. These gender differences warrant further study and may affect prognosis, follow-up, and timing of valve surgery., (Copyright © 2019 Australian and New Zealand Society of Cardiac and Thoracic Surgeons (ANZSCTS) and the Cardiac Society of Australia and New Zealand (CSANZ). Published by Elsevier B.V. All rights reserved.)

Background: High resolution 2D whole slide imaging provides rich information about the tissue structure. This information can be a lot richer if these 2D images can be stacked into a 3D tissue volume. A 3D analysis, however, requires accurate reconstruction of the tissue volume from the 2D image stack. This task is not trivial due to the distortions such as tissue tearing, folding and missing at each slide. Performing registration for the whole tissue slices may be adversely affected by distorted tissue regions. Consequently, regional registration is found to be more effective. In this paper, we propose a new approach to an accurate and robust registration of regions of interest for whole slide images. We introduce the idea of multi-scale attention for registration., Results: Using mean similarity index as the metric, the proposed algorithm (mean ± SD [Formula: see text]) followed by a fine registration algorithm ([Formula: see text]) outperformed the state-of-the-art linear whole tissue registration algorithm ([Formula: see text]) and the regional version of this algorithm ([Formula: see text]). The proposed algorithm also outperforms the state-of-the-art nonlinear registration algorithm (original: [Formula: see text], regional: [Formula: see text]) for whole slide images and a recently proposed patch-based registration algorithm (patch size 256: [Formula: see text] , patch size 512: [Formula: see text]) for medical images., Conclusion: Using multi-scale attention mechanism leads to a more robust and accurate solution to the problem of regional registration of whole slide images corrupted in some parts by major histological artifacts in the imaged tissue.

Purpose: The purpose of this study was to compare the outcomes of arthroscopic rotator cuff repair (ARCR) in patients with preoperative stiffness to those without., Methods: A total of 135 patients were prospectively evaluated for 2 years after ARCR for small to medium sized rotator cuff tears at our institution. Patients were divided into stiff (<100° of passive forward flexion) and non-stiff cohorts. The stiff group underwent manipulation under anesthesia (MUA) before ARCR was performed. Outcomes were measured using visual analog scale (VAS), Constant Shoulder Score (CSS), and Oxford Shoulder Score (OSS) recorded at the preoperative, 6- , 12-, and 24-month time points. The results of ARCR between the cohorts were then compared., Results: A total of 123 out of 135 patients (91.1%) completed the follow-up (stiff n = 46, non-stiff n = 77). There were significant improvements in the mean CSS scores at 6 (mean, 59.87; P < .001) and 12 months (mean, 65.88; P = .021) in the stiff group. There were no significant differences detectable in the CSS and OSS scores between the stiff and non-stiff groups at 6, 12, and 24 months. However, the percentage of patients achieving minimal clinically important difference was significantly higher in the stiff group (97.8%) compared with the non-stiff group (75.3%; P = .001). The VAS scores, forward flexion, and strength in both groups were found to be comparable., Conclusions: The results of our study showed no significant differences in outcomes scores in patients with stiff shoulders who underwent MUA combined with ARCR compared with patients with non-stiff shoulders who underwent ARCR alone. Therefore, early surgical repair should be considered in patients with rotator cuff tears and concomitant shoulder stiffness., Level of Evidence: Level II, prospective cohort study., (Copyright © 2020 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)

Biliary tract cancer (BTC) is a rare, but aggressive, disease that comprises of gallbladder carcinoma, intrahepatic cholangiocarcinoma and extrahepatic cholangiocarcinoma, with heterogeneous molecular profiles. Advanced disease has limited therapeutic options beyond first-line platinum-based chemotherapy. Immunotherapy has emerged as a viable option for many cancers with a similar unmet need. Therefore, we reviewed current understanding of the tumor immune microenvironment and recent advances in cellular immunotherapy and therapeutic cancer vaccines against BTC. We illustrated the efficacy of dendritic cell vaccination in one patient with advanced, chemorefractory, melanoma-associated antigen (MAGE)-positive gallbladder carcinoma, who was given multiple injections of an allogenic MAGE antigen-positive melanoma cell lysate (MCL)-based autologous dendritic cell vaccine combined with sequential anti-angiogenic therapy. This resulted in good radiological and tumor marker response and an overall survival of 3 years from diagnosis. We postulate the potential synergism of adding anti-angiogenic therapy, such as bevacizumab, to immunotherapy in BTC, as a rational scientific principle to positively modulate the tumor microenvironment to augment antitumor immunity.

This work reports on the structural and photophysical characterization of D 4 -symmetrical octanuclear circular Ln III helicates, [( R )- or ( S )- i Pr-Pybox] 8 (Ln III ) 8 (THP) 8 (where Ln = Eu and Tb, THP = trianionic tris-β-diketonate, and i Pr-Pybox = chiral bis(4-isopropyl-2-oxazolinyl)pyridine). X-ray crystallographic analysis revealed that the octanuclear circular helicate possesses square antiprism architecture and consists of four [( R )- or ( S )- i Pr-Pybox] 2 Ln III 2 (THP) 2 asymmetric units arranged in a closed ring form. Ligand-to-ligand interactions between the THP and the i Pr-Pybox ligands have successfully directed formation of enantiopure, homoconfigurational (Δ,Δ,Δ,Δ,Δ,Δ,Δ,Δ)- R and (Λ,Λ,Λ,Λ,Λ,Λ,Λ,Λ)- S isomers. All of the nonacoordinated Ln III ions are identical and exhibit a distorted capped square antiprism (CSAP) geometry. Upon excitation of the ligand absorption band (λ = 360 nm), the circular helicates display characteristic Eu III (red, 5 D 0 → 7 F J , J = 0-4) or Tb III (green, 5 D 4 → 7 F J , J = 6-3) core f-f luminescence. The overall emission quantum yields of the circular Eu III and Tb III helicates are 0.145 and 0.0013, respectively, in chloroform. The Eu III and Tb III complexes exhibit remarkable circularly polarized luminescence (CPL) activity at their magnetic dipole transition with observed luminescence dissymmetry factors | g lum | of 1.25 ( 5 D 0 → 7 F 1 , λ = 592 nm) and 0.25 ( 5 D 4 → 7 F 5, λ = 541 nm), respectively. Exceptional | g lum | values of the circular Eu III helicates highlight the visible intensity difference between left and right circularly polarized emissions of R and S isomers in chloroform and PMMA thin film.

Three new tetranuclear europium(III) alternating circular helicates, ( 2- Ph RRRR/ 2- Ph SSSS ) [( R )- or ( S )-Ph-Pybox] 4 Eu III 4 (BPP) 6 , ( 2- iPr RRRR / 2- iPr SSSS ) [( R )- or ( S )- i Pr-Pybox] 4 Eu III 4 (BPP) 6 , and ( 3- Ph RRRR/ 3- Ph SSSS ) [( R )- or ( S )-Ph-Pybox] 4 Eu III 4 (BHP) 6 , are presented with their structural and chiroptical characterization (BPP and BHP = bis-β-diketonates; Pybox = chiral bis(oxazolinyl) pyridine ligand). X-ray crystallographic analysis revealed that different extents of interligand-contacting interactions between bis-β-diketonates and Pybox ligands produce different bis-β-diketonates arrangements around four Eu III ions and, thus, their specific symmetry in the final tetranuclear complexes. 2- Ph RRRR/ 2- Ph SSSS and 3- Ph RRRR/ 3- Ph SSSS are recognized as D 2 -symmetry, while 2- iPr RRRR / 2- iPr SSSS self-assemblies possess pseudo C 2h -symmetry. Due to different molecular symmetry, 2- Ph RRRR/ 2- Ph SSSS and 2- iPr RRRR / 2- iPr SSSS crystals display different ligands orientation in their Eu III coordination spheres. The presence of pseudo σ h -mirror symmetry in 2- iPr RRRR / 2- iPr SSSS promotes a pair of distinguishable Eu III geometries. All the chiral self-assemblies exhibit almost identical photoluminescence emission spectra patterns of f-f transitions in the Eu III core. The Eu III self-assemblies exhibit intense CPL with different observed | g lum | values ( 2- Ph RRRR/ 2- Ph SSSS ∼ |0.31|, 2- iPr RRRR / 2- iPr SSSS ∼ |0.08|, 3- Ph RRRR/ 3- Ph SSSS ∼ |0.31| in chloroform).

Background: H7N9 avian influenza is an infection of public health concern, in part because of its high mortality rate and pandemic potential., Aims: To describe the clinical features of H7N9 avian influenza and the response to treatment., Methods: Clinical, radiological and histopathological data, and treatment-related of H7N9-infected patients hospitalised during 2014-2017 were extracted and analysed., Results: A total of 17 H7N9 patients (three females; mean age, 58.4 ± 13.7 years) was identified; of these six died. All patients presented with fever and productive cough; four patients had haemoptysis and 13 had chest distress and/or shortness of breath. Early subnormal white blood cell count and elevation of serum liver enzymes were common. Multilobar patchy shadows, rapid progression to ground-glass opacities, air bronchograms and consolidation were the most common imaging findings. Histopathological examination of lung tissue of three patients who died showed severe alveolar epithelial cell damage, with inflammatory exudation into the alveolar space and hyaline membrane formation; widened alveolar septae, prominent inflammatory cell infiltration; and hyperplasia of pneumocytes. Viral inclusions were found in the lung tissue of two patients. All patients received antiviral drugs (oseltamivir ± peramivir). Four patients carried the rs12252-C/C interferon-induced transmembrane protein-3 (IFITM3) genotype, while the others had the C/T genotype., Conclusions: H7N9 virus infection causes human influenza-like symptoms, but may rapidly progress to severe pneumonia and even death. Clinicians should be alert to the possibility of H7N9 infection in high-risk patients. The presence of the IFITM3 rs12252-C genotype may predict severe illness., (© 2019 Royal Australasian College of Physicians.)

Highly emissive fluorophores based on polyaromatic hydrocarbons with tunable emission properties and aggregated structures play a very important role in relevant functional studies. In this study, a novel alkynylpyrene derivative 1 was synthesized, which exhibits unimolecular to excimer emission in methanol with an increasing concentration accompanied by the formation of nanovesicles via the π-π stacking, hydrogen bond and hydrophobic interaction. The self-assembly behavior as well as emission properties of 1 in aprotic polar solvents (ACN, acetone, DMF and DMSO) can also be adjusted by the volume fraction of the poor solvent H2O, which can induce 1 self-assembly to excimer state and could be applied in information transfer. Moreover, upon visible light irradiation, photoswitchable performance of nanovesicles of 1 was observed in which the emission markedly changes from yellow to blue; this is attributed to the cycloaddition reaction of alkynyl groups and singlet oxygen, which can be generated without the addition of external photosensitizers. The multi-responsive and fluorescence behavior of the alkynylpyrene derivative show that the self-assembly can be used to expand the development of this type of fluorophores, and the novel photoinduced tunability of the fluorescence emission provides an effective strategy to obtain high-performance transmitting and sensing materials.

Background: Recent reports suggest that component plasma products contain significant quantities of cellular contamination. We hypothesized that leukoreduction of whole blood before preparation of derived plasma is an effective method to prevent cellular contamination of stored plasma., Study Design: Samples of never-frozen liquid plasma prepared by standard methods (n = 25) were obtained from 3 regional blood centers that supply 3 major trauma centers. Samples were analyzed for leukocyte and platelet contamination by flow cytometry. To determine if leukoreduction of whole blood before centrifugation and expression of plasma prevents cellular contamination of liquid plasma, 1 site generated 6 additional units of liquid plasma from leukoreduced whole blood, which were then compared with units of liquid plasma derived by standard processing., Results: Across all centers, each unit of never-frozen liquid plasma contained a mean of 12.8 ± 3.0 million leukocytes and a mean of 4.6 ± 2 billion platelets. Introduction of whole blood leukoreduction (LR) before centrifugation and plasma extraction essentially eliminated all contaminating leukocytes (Non-LR: 12.3 ± 2.9 million vs LR: 0.05 ± 0.05 million leukocytes) and platelets (Non-LR: 4.2 ± 0.3 billion platelets vs LR: 0.00 ± 0.00 billion platelets)., Conclusions: Despite widespread belief that stored plasma is functionally acellular, testing of liquid plasma from 3 regional blood banks revealed a significant amount of previously unrecognized cellular contamination. Introduction of a leukoreduction step before whole blood centrifugation essentially eliminated detectable leukocyte and platelet contaminants from plasma. Therefore, our study highlights a straightforward and cost-effective method to eliminate cellular contamination of stored plasma., (Copyright © 2020. Published by Elsevier Inc.)

C-reactive protein (CRP) is a major human acute-phase reactant that is composed of five identical subunits. CRP dissociates into subunits at inflammatory loci forming monomeric CRP (mCRP) with substantially enhanced activities, which can be further activated by reducing the intra-subunit disulfide bond. However, conformational changes underlying the activation process of CRP are less well understood. Conformational changes accompanying the conversion of CRP to mCRP with or without reduction were examined with circular dichroism spectroscopy, fluorescence spectroscopy, electron microscopy, size-exclusion chromatography, and neoepitope expression. The conversion of CRP to mCRP follows a two-stage process. In the first stage, CRP dissociates into molten globular subunits characterized by intact secondary structure elements with greatly impaired tertiary packing. In the second stage, these intermediates completely lose their native subunit conformation and assemble into high-order aggregates. The inclusion of reductant accelerates the formation of molten globular subunits in the first step and promotes the formation of more compact aggregates in the second stage. We further show a significant contribution of electrostatic interactions to the stabilization of native CRP. The conformational features of dissociated subunits and the aggregation of mCRP may have a key impact on their activities.

The H 2 O/D 2 O/HDO isotopomers of water are presented in terms that enable bond-flexing, bond-twist and bond-anharmonicity to be quantified during the bending (Q1), symmetric-stretch (Q2) and anti-symmetric-stretch (Q3) normal modes of vibration. Bond-flexing was detected by the presence of curved bonding and the bond-anharmonicity was detected by the presence of motion of the bond critical point (BCP) relative to the oxygen atom. These 2-D scalar measures are unable to fully describe the 3-D nature of the normal modes of vibration and therefore any susceptibility towards normal mode coupling, or to fully distinguish the three isotopomers. To detect bond-twist a vector-based measure was used, in the form of the bond critical point (BCP) trajectory, constructed in terms of preferred directions of electronic motion, defined by the variation of the position of the BCP during the normal modes of vibration. The BCP trajectories describe the coupling of the intramolecular bending and symmetric-stretch normal modes as well as distinguishing all three isotopomers within the harmonic approximation. The coupling of the bending and symmetric-stretch normal modes are suggested to be facilitated by the absence of bond-twist that would disrupt the coupling between Sigma O-H bonds and hydrogen-bonding. Partial coupling was found for the mixed isotopomer HDO.

Background: Transplantation of lungs procured after donation after circulatory death (DCD) is challenging because postmortem metabolic degradation may engender susceptibility to ischemia-reperfusion (IR) injury. Because oxidative mitochondrial DNA (mtDNA) damage has been linked to endothelial barrier disruption in other models of IR injury, here we used a fusion protein construct targeting the DNA repair 8-oxoguanine DNA glycosylase-1 (OGG1) to mitochondria (mtOGG1) to determine if enhanced repair of mtDNA damage attenuates endothelial barrier dysfunction after IR injury in a rat model of lung procurement after DCD., Materials and Methods: Lungs excised from donor rats 1 h after cardiac death were cold stored for 2 h after which they were perfused ex vivo in the absence and presence of mt-OGG1 or an inactive mt-OGG1 mutant. Lung endothelial barrier function and mtDNA integrity were determined during and at the end of perfusion, respectively., Results and Conclusions: Mitochondria-targeted OGG1 attenuated indices of lung endothelial dysfunction incurred after a 1h post-mortem period. Oxidative lung tissue mtDNA damage as well as accumulation of proinflammatory mtDNA fragments in lung perfusate, but not nuclear DNA fragments, also were reduced by mitochondria-targeted OGG1. A repair-deficient mt-OGG1 mutant failed to protect lungs from the adverse effects of DCD procurement., Conclusions: These findings suggest that endothelial barrier dysfunction in lungs procured after DCD is driven by mtDNA damage and point to strategies to enhance mtDNA repair in concert with EVLP as a means of alleviating DCD-related lung IR injury., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Stored plasma products are widely regarded as being functionally acellular, obviating the need for leukoreduction. We tested the hypothesis that donor plasma is contaminated by leukocytes and platelets, which, after frozen storage, would release cellular debris in quantities sufficient to elicit significant pro-inflammatory responses., Study Design: Samples of never-frozen liquid plasma from 2 regional Level I trauma centers were analyzed for leukocyte and platelet contamination. To determine if the cellular contamination and associated debris found in liquid plasma were at levels sufficient to evoke an innate immune response, known quantities of leukocytes were subjected to a freeze-thaw cycle, added to whole blood, and the magnitude of the inflammatory response was determined by induction of interleukin-6., Results: Units of never-frozen plasma from 2 regional Level I trauma centers located in Alabama and Louisiana contained significant amounts of leukocyte contamination (Louisiana, n = 22; 17.3 ± 4.5 million vs Alabama, n = 22; 11.3 ± 2.2 million) and platelet contamination (Louisiana, n = 21; 0.86 ± 0.20 billion vs Alabama, n = 22; 1.0 ± 0.3 billion). Cellular debris from as few as 1 million leukocytes induced significant increases in interleukin-6 levels (R 2  = 0.74; p < 0.0001)., Conclusions: Stored plasma units from trauma center blood banks were highly contaminated with leukocytes and platelets, at levels more than 15-fold higher than sufficient to elicit ex vivo inflammatory responses. In light of paradigm shifts toward the use of more empiric plasma for treatment of hypovolemia, this study suggests that new manufacturing and quality-control processes are needed to eliminate previously unrecognized cellular contamination present in stored plasma products., (Copyright © 2019 American College of Surgeons. Published by Elsevier Inc. All rights reserved.)

Three typical plant communities (evergreen broad-leaved forest at low-altitude 1100 m, evergreen and deciduous mixed broad-leaved forest at mid-altitude 1500 m, and evergreen conife-rous and broad-leaved mixed forest at high-altitude 1900 m) in Maoer Mountain, Guangxi, China were surveyed along an altitude gradient. We measured the tree layer plant architecture and environmental factors, to analyze the variation of plant architecture traits among the three communities and its influencing factors. The results showed that the tree layer canopy area, basal diameter at 45 cm height, diameter at breast height (DBH), and leaf convergence increased with increasing altitude, whereas tree height, branch height, and canopy thickness first increased and then decreased. Horizontal branches occurred more often in communities at lower altitude , less frequent at high altitude, and the least frequent in middle altitude communities. Correlations among tree layer plant architecture traits were stronger in the mid-altitude community than that in the other altitude communities. Results from the redundancy analysis showed that soil organic matter and total solar radiation were the main factors driving the variation of plant architecture traits in the tree layers, accounting for 39.6% and 23.9% of the total variation, respectively. Soil organic matter had a greater positive impact on canopy area and branch height, whereas total solar radiation was more influential on the DBH and 45 cm basal diameter. In conclusion, tree layer architecture of communities along the altitude gradient in Maoer Mountain was divergent, with soil organic matter and total solar radiation as the main driving forces.

Rationale: Neuroblastoma is one of the most common malignant tumors in childhood, which mainly occurs in adrenal glands and peripheral sympathetic nerve system. Neuroblastoma occurring in adulthood is rare, and adults with neuroblastoma arising from thorax are exceedingly rare. A case of neuroblastoma that originated from thorax was reported, and was treated by resection operation., Patient Concerns: A 46-year-old woman was admitted to our hospital with left side chest pain for 5 days. Laboratory examinations were all normal. Chest computerized tomogram (CT) showed a lesion with clear boundary that was located at the left dorsal pleura. The nature of the mass was heterogeneous, showing slight heterogeneous enhancement after contrast and there was no obvious necrosis., Diagnoses: Based on the morphologic and immunohistochemical features, the tumor diagnosis was favorable for neuroblastoma., Interventions: A resection operation was carried out., Outcomes: Three years postoperative, no sign of recurrence or metastasis has been observed., Lessons: Primary neuroblastoma in adulthood is rare and has poor prognosis. Resection can be an important treatment option, and combining with other methods like chemotherapy, stem cell transplantation, the survival rate may be improved.

Chikungunya virus (CHIKV) is transmitted to humans through mosquitoes and causes Chikungunya fever. Nonstructural protein 2 (nsP2) exhibits the protease and RNA helicase activities that are required for viral RNA replication and transcription. Unlike for the C-terminal protease, the structure of the N-terminal RNA helicase (nsP2h) has not been determined. Here, we report the crystal structure of the nsP2h bound to the conserved 3'-end 14 nucleotides of the CHIKV genome and the nonhydrolyzable transition-state nucleotide analog ADP-AlF 4 Overall, the structural analysis revealed that nsP2h adopts a uniquely folded N-terminal domain followed by a superfamily 1 RNA helicase fold. The conserved helicase motifs establish polar contacts with the RNA backbone. There are three hydrophobic residues (Y161, F164, and F287) which form stacking interactions with RNA bases and thereby bend the RNA backbone. An F287A substitution that disrupted these stacking interactions increased the basal ATPase activity but decreased the RNA binding affinity. Furthermore, the F287A substitution reduced viral infectivity by attenuating subgenomic RNA synthesis. Replication of the mutant virus was restored by pseudoreversion (A287V) or adaptive mutations in the RecA2 helicase domain (T358S or V410I). Y161A and/or F164A substitutions, which were designed to disrupt the interactions with the RNA molecule, did not affect the ATPase activity but completely abolished the replication and transcription of viral RNA and the infectivity of CHIKV. Our study sheds light on the roles of the RNA helicase region in viral replication and provides insights that might be applicable to alphaviruses and other RNA viruses in general., Competing Interests: The authors declare no conflict of interest.

Background: Previous data suggest that the immune microenvironment plays a critical role in human epidermal growth factor receptor 2 (HER2) -positive breast cancer; however, there is little known about the immune profiles of small HER2-positive tumors. In this study, we aimed to characterize the immune microenvironment of small HER2-positive breast cancers included in the Adjuvant paclitaxel and trastuzumab for node-negative, HER2-positive breast cancer (APT) trial and to correlate the immune markers with pathological and molecular tumor characteristics., Patients and Methods: The APT trial was a multicenter, single-arm, phase II study of paclitaxel and trastuzumab in patients with node-negative HER2-positive breast cancer. The study included 406 patients with HER2-positive, node-negative breast cancer, measuring up to 3 cm. Exploratory analysis of tumor infiltrating lymphocytes (TIL), programmed death-ligand 1 (PD-L1) expression (by immunohistochemistry), and immune gene signatures using data generated by nCounter PanCancer Pathways Panel (NanoString Technologies, Seattle, WA), and their association with pathological and molecular characteristics was carried out., Results: Of the 406 patients, 328 (81%) had at least one immune assay carried out: 284 cases were evaluated for TIL, 266 for PD-L1, and 213 for immune gene signatures. High TIL (≥60%) were seen with greater frequency in hormone-receptor (HR) negative, histological grades 2 and 3, as well in HER2-enriched and basal-like tumors. Lower stromal PD-L1 (≤1%) expression was seen with greater frequency in HR-positive, histological grade 1, and in luminal tumors. Both TIL and stromal PD-L1 were positively correlated with 10 immune cell signatures, including Th1 and B cell signatures. Luminal B tumors were negatively correlated with those signatures. Significant correlation was seen among these immune markers; however, the magnitude of correlation did not indicate a monotonic relationship between them., Conclusion: Immune profiles of small HER2-positive breast cancers differ according to HR status, histological grade, and molecular subtype. Further work is needed to explore the implication of these findings on disease outcome., Clinical Trial Registration: clinicaltrials.gov identifier: NCT00542451., (© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)